1、Designation: D7600 091Standard Test Method forDetermination of Aldicarb, Carbofuran, Oxamyl andMethomyl by Liquid Chromatography/Tandem MassSpectrometry1This standard is issued under the fixed designation D7600; the number immediately following the designation indicates the year oforiginal adoption
2、or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. Asuperscript epsilon () indicates an editorial change since the last revision or reapproval.1NOTEThis Test Method was changed editorially in 2012.1. Scope1.1 This procedure covers t
3、he determination of aldicarb,carbofuran, oxamyl and methomyl (referred to collectively ascarbamates in this test method) in surface water by directinjection using liquid chromatography (LC) and detected withtandem mass spectrometry (MS/MS). These analytes are quali-tatively and quantitatively determ
4、ined by this method. Thismethod adheres to multiple reaction monitoring (MRM) massspectrometry.1.2 This test method has been developed by US EPARegion5 Chicago Regional Laboratory (CRL).1.3 The values stated in SI units are to be regarded asstandard. No other units of measurement are included in thi
5、sstandard.1.4 The Detection Verification Level (DVL) and ReportingRange for the carbamates are listed in Table 1.1.4.1 The DVL is required to be at a concentration at least3 times below the Reporting Limit (RL) and have a signal/noise ratio greater than 3:1. Fig. 1 displays the signal/noiseratios of
6、 the primary single reaction monitoring (SRM) transi-tions and Fig. 2 displays the confirmatory SRM transitions atthe DVLs for the carbamates.1.4.2 The reporting limit is the concentration of the Level 1calibration standard as shown in Table 2 for the carbamates.1.5 This standard does not purport to
7、 address all of thesafety concerns, if any, associated with its use. It is theresponsibility of the user of this standard to establish appro-priate safety and health practices and determine the applica-bility of regulatory limitations prior to use.2. Referenced Documents2.1 ASTM Standards:2D1129 Ter
8、minology Relating to WaterD1193 Specification for Reagent WaterD2777 Practice for Determination of Precision and Bias ofApplicable Test Methods of Committee D19 on WaterD3856 Guide for Management Systems in LaboratoriesEngaged in Analysis of WaterD3694 Practices for Preparation of Sample Containers
9、andfor Preservation of Organic ConstituentsD5847 Practice for Writing Quality Control Specificationsfor Standard Test Methods for Water AnalysisE2554 Practice for Estimating and Monitoring the Uncer-tainty of Test Results of a Test Method in a SingleLaboratory Using a Control Sample Program2.2 Other
10、 Documents:EPApublication SW-846 Test Methods for Evaluating SolidWaste, Physical/Chemical Methods33. Terminology3.1 Definitions:3.1.1 detection verification level (DVL), na concentrationthat has a signal/noise ratio greater than 3:1 and is at least 3times below the reporting limit (RL).3.1.2 report
11、ing limit (RL), nthe concentration of thelowest-level calibration standard used for quantification.3.1.3 carbamates, nin this test method, aldicarb, carbo-furan, oxamyl and methomyl collectively.1This test method is under the jurisdiction of ASTM Committee D19 on Waterand is the direct responsibilit
12、y of Subcommittee D19.06 on Methods forAnalysis forOrganic Substances in Water.Current edition approved Dec. 1, 2009. Published February 2012. DOI: 10.1520/D7600-09E01.2For referenced ASTM standards, visit the ASTM website, www.astm.org, orcontact ASTM Customer Service at serviceastm.org. For Annual
13、 Book of ASTMStandards volume information, refer to the standards Document Summary page onthe ASTM website.3Available from National Technical Information Service (NTIS), U.S. Depart-ment of Commerce, 5285 Port Royal Road, Springfield, VA, 22161 or at http:/www.epa.gov/epawaste/hazard/testmethods/ind
14、ex.htm.1Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.3.2 Abbreviations:3.2.1 pptparts per trillion, ng/L3.2.2 NDnon-detect4. Summary of Test Methods4.1 This is a performance-based method and modificationsare allowed to improve perf
15、ormance.4.2 For carbamate analysis, samples are shipped to the labbetween 0C and 6C and analyzed within 7 days of collection.In the lab, the samples are spiked with surrogate, filtered usinga syringe driven Millex HV PVDF filter unit and analyzeddirectly by LC/MS/MS.4.3 Aldicarb, carbofuran, oxamyl,
16、 methomyl and 4-bromo-3,5-dimethylphenyl-N-methylcarbamate (BDMC, surrogate)are identified by retention time and two SRM transitions. Thetarget analytes and surrogate are quantitated using the primarySRM transitions utilizing an external calibration. The finalreport issued for each sample lists the
17、concentration of aldi-carb, carbofuran, oxamyl, methomyl and the BDMC surrogaterecovery.5. Significance and Use5.1 The N-methyl carbamate (NMC) pesticides: aldicarb,carbaryl, carbofuran, formetanate hydrochloride, methiocarb,methomyl, oxamyl, pirimicarb, propoxur, and thiodicarb havebeen identified
18、by EPA as working through a common mecha-nism. They affect the nervous system by reducing the ability ofthe enzyme cholinesterase. Cholinesterase inhibition was theprimary toxicological effect of regulatory concern to EPA inassessing the NMCs food, drinking water and residential risks.In most of the
19、 country, NMC residues in drinking watersources are at levels that are not likely to contribute substan-tially to the multi-pathway cumulative exposure. Shallowprivate wells extending through highly permeable soils intoshallow, acidic ground water represent what the EPA believesto be the most vulner
20、able drinking water.45.2 This method has been investigated for use with reagentand surface water for the selected carbamates: aldicarb, carbo-furan, oxamyl and methomyl.6. Interferences6.1 Method interferences may be caused by contaminants insolvents, reagents, glassware and other apparatus producin
21、gdiscrete artifacts or elevated baselines. All of these materialsare demonstrated to be free from interferences by analyzinglaboratory reagent blanks under the same conditions assamples.6.2 All glassware is washed in hot water with a detergent,rinsed in hot water followed by distilled water. The gla
22、sswareis then dried and heated in an oven at 250C for 15 to 30minutes. All glassware is subsequently cleaned with acetone,then methanol.6.3 All reagents and solvents should be pesticide residuepurity or higher to minimize interference problems.6.4 Matrix interferences may be caused by contaminantsth
23、at are co-extracted from the sample. The extent of matrixinterferences can vary considerably from sample source de-pending on variations of the sample matrix.7. Apparatus7.1 LC/MS/MS System7.1.1 Liquid Chromatography (LC) SystemA completeLC system is needed in order to analyze samples.5A systemthat
24、is capable of performing at the flows, pressures, controlledtemperatures, sample volumes and requirements of the stan-dard may be used.7.1.2 Analytical Column-WatersXBridge C18, 150 mm 32.1 mm, 3.5 m particle size, or equivalent.7.1.3 Tandem Mass Spectrometer (MS/MS) SystemAMS/MS system capable of M
25、RM analysis.6A system that iscapable of performing at the requirements in this standard maybe used.7.2 Filtration Device7.2.1 Hypodermic syringeAlock tip glass syringe capableof holding a Millex HV Syringe Driven Filter Unit PVDF 0.45m (Millipore Corporation, Catalog # SLHV033NS) or similarmay be us
26、ed.7.2.1.1 A 25-mL lock tip glass syringe size is recommendedsince a 25-mL sample size is used in this test method.7.2.2 FilterMillex HV Syringe Driven Filter Unit PVDF0.45 m (Millipore Corporation, Catalog # SLHV033NS) orsimilar may be used.8. Reagents and Materials8.1 Purity of ReagentsHigh-perfor
27、mance liquid chroma-tography (HPLC) pesticide residue analysis and spectropho-tometry grade chemicals shall be used in all tests. Unlessindicated otherwise, it is intended that all reagents shallconform to the Committee on Analytical Reagents of theAmerican Chemical Society.7Other reagent grades may
28、 beused provided they are first determined to be of sufficientlyhigh purity to permit their use without affecting the accuracy ofthe measurements.8.2 Purity of WaterUnless otherwise indicated, referencesto water shall be understood to mean reagent water conforming4Additional information about carbam
29、ate pesticides can be found on the Internetat http:/www.epa.gov (2009).5A Waters Alliance High Performance Liquid Chromatography (HPLC) Systemwas used to develop this test method. The multi-laboratory study included Agilent,Thermo Electron and Waters LC systems.6A Waters Quattro micro API mass spect
30、rometer was used to develop this testmethod. The multi-laboratory study included Agilent, Applied Biosystems, ThermoElectron, Varian and Waters mass spectrometers.7Reagent Chemicals, American Chemical Society Specifications, AmericanChemical Society, Washington, DC. For Suggestions on the testing of
31、 reagents notlisted by the American Chemical Society, see Annual Standards for LaboratoryChemicals, BDH Ltd., Poole, Dorset, U.K., and the United States Pharmacopeiaand National Formulary, U.S. Pharmacopeial Convention, Inc. (USPC), Rockville,MD.TABLE 1 Detection Verification Level and Reporting Ran
32、geAnalyte DVL (g/L) Reporting Range (g/L)Aldicarb 100 1100Carbofuran 100 1100Oxamyl 100 1100Methomyl 100 1100D7600 0912FIG. 1 Example Primary SRM Chromatograms Signal/Noise RatiosFIG. 2 Example Confirmatory SRM Chromatograms Signal/Noise RatiosD7600 0913to Type 1 of Specification D1193. It must be d
33、emonstrated thatthis water does not contain contaminants at concentrationssufficient to interfere with the analysis.8.3 GasesUltrapure nitrogen and argon.8.4 Acetonitrile (CAS # 75-05-8).8.5 Methanol (CAS # 67-56-1).8.6 Acetone (CAS # 67-64-1).8.7 Ammonium acetate (CAS # 631-61-8).8.8 Ammonium hydro
34、xide (Concentrated, CAS # 1336-21-6).8.9 Aldicarb (CAS # 116-06-3).8.10 Carbofuran (CAS # 1563-66-2).8.11 Oxamyl (CAS # 23135-22-0).8.12 Methomyl (CAS # 16752-77-5).8.13 4-Bromo-3,5-dimethylphenyl-N-methylcarbamate(BDMC, CAS # 672-99-1).8.13.1 BDMC is used as a surrogate in this standard.9. Hazards9
35、.1 Normal laboratory safety applies to this method. Ana-lysts should wear safety glasses, gloves, and lab coats whenworking in the lab. Analysts should review the Material SafetyData Sheets (MSDS) for all reagents used in this method.10. Sampling10.1 SamplingGrab samples must be collected in$25-mL p
36、re-cleaned amber glass bottles with Teflon-linedcaps demonstrated to be free of interferences. This test methodrequires a 25-mL sample size per analysis. Conventionalsampling practices should be followed. Refer to Guide D3856and Practices D3694.10.2 PreservationStore samples between 0C and 6Cfrom th
37、e time of collection until analysis. Analyze the samplewithin 7 days of collection.11. Preparation of LC/MS/MS11.1 LC Chromatograph Operating Conditions5:11.1.1 Injection volumes of all calibration standards andsamples are 100 L. The first sample analyzed after thecalibration curve is a blank to ens
38、ure there is no carry-over. Thegradient conditions for the liquid chromatograph are shown inTable 3.11.1.2 TemperaturesColumn, 30C; Sample compart-ment, 15C.11.1.3 Seal WashSolvent: 50 % Acetonitrile/50 % Water;Time: 5 minutes.11.1.4 Needle WashSolvent: 50 % Acetonitrile/50 % Wa-ter; Normal wash, ap
39、proximately 13 second wash time.11.1.5 Autosampler PurgeThree loop volumes.11.1.6 Specific instrument manufacturer wash/purge speci-fications should be followed in order to eliminate samplecarry-over in the analysis of carbamates.11.2 Mass Spectrometer Parameters6:11.2.1 In order to acquire the maxi
40、mum number of datapoints per SRM channel while maintaining adequate sensitiv-ity, the tune parameters may be optimized according to yourinstrument. Each peak requires at least 10 scans per peak foradequate quantitation. This standard contains only one surro-gate and four target compounds. The MRM ex
41、periment win-dows were set to acquire methomyl and oxamyl in oneexperiment window while aldicarb, carbofuran and BDMC arein their individual MRM experiment windows. This is requiredbecause the chromatographic resolution separating oxamyl andmethomyl was not achieved. Variable parameters regardingret
42、ention times, SRM Transitions and cone and collisionenergies are shown in Table 4.The instrument is set in the Electrospray (+) positive setting.Capillary Voltage: 3.5 kVCone: Variable depending on analyte (Table 4)Extractor: 2 VoltsRF Lens: 0.2 VoltsSource Temperature: 120CDesolvation Temperature:
43、300CDesolvation Gas Flow: 500 L/hrCone Gas Flow: 25 L/hrLow Mass Resolution 1: 14.5High Mass Resolution 1: 14.5Ion Energy 1: 0.5Entrance Energy: 1Collision Energy: Variable depending on analyte (Table 4)Exit Energy: 2Low Mass Resolution 2: 15High Mass resolution 2: 15Ion Energy 2: 0.5Multiplier: 650
44、Gas Cell Pirani Gauge: 3.3 3 103TorrInter-Channel Delay: 0.02 secondsInter-Scan Delay: 0.1 secondsRepeats: 1Span: 0 DaltonsDwell: 0.1 Seconds12. Calibration and Standardization12.1 The mass spectrometer must be calibrated per manu-facturer specifications before analysis. In order that analyticalvalu
45、es obtained using this test method are valid and accuratewithin the confidence limits of the test method, the followingprocedures must be followed when performing the test method.TABLE 2 Concentrations of Calibration Standards (PPB)Analyte/Surrogate LV 1 LV 2 LV 3 LV 4 LV 5 LV 6Aldicarb 1 5 10 25 50
46、 100Carbofuran 1 5 10 25 50 100Oxamyl 1 5 10 25 50 100Methomyl 1 5 10 25 50 100BDMC (Surrogate) 2 10 20 50 100 200TABLE 3 Gradient Conditions for Liquid ChromatographyTime(min)Flow(L/min)PercentCH3CNPercent 95 %Water/ 5 %CH3CNPercent50 mmolarNH4OAc/NH4OHin 95 % Water/5 %CH3CN0 300 0 95 52 300 0 95 5
47、4 300 30 65 56 300 35 60 58 300 35 60 510 300 75 20 511.5 300 75 20 512 300 95 0 518 300 95 0 520 300 0 95 523 300 0 95 5D7600 091412.2 Calibration and StandardizationTo calibrate the in-strument, analyze six calibration standards containing the sixconcentration levels of the carbamates and BDMC sur
48、rogateprior to analysis as shown in Table 2. A calibration stockstandard solution is prepared from standard materials orpurchased as certified solutions. Stock standard solution A(Level 6) containing aldicarb, carbofuran, oxamyl, methomyland BDMC is prepared at Level 6 concentration and aliquots oft
49、hat solution are diluted to prepare Levels 1 through 5. Thefollowing steps will produce standards with the concentrationvalues shown in Table 2. The analyst is responsible forrecording initial component weights carefully when workingwith pure materials and correctly carrying the weights throughthe dilution calculations.12.2.1 Prepare stock standard solution A (Level 6) byadding to a 100-mL volumetric flask individual methanolsolutions of the following: 50 L of aldicarb, carbofuran,oxamyl and methomyl each at 0.2 g/L and 50 L of
